CN108892803A - A kind of salt tolerant anion-exchange chromatography medium and preparation method thereof - Google Patents
A kind of salt tolerant anion-exchange chromatography medium and preparation method thereof Download PDFInfo
- Publication number
- CN108892803A CN108892803A CN201810963586.XA CN201810963586A CN108892803A CN 108892803 A CN108892803 A CN 108892803A CN 201810963586 A CN201810963586 A CN 201810963586A CN 108892803 A CN108892803 A CN 108892803A
- Authority
- CN
- China
- Prior art keywords
- salt tolerant
- exchange chromatography
- chromatography medium
- polyamines
- anion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000012501 chromatography medium Substances 0.000 title claims abstract description 66
- 150000003839 salts Chemical class 0.000 title claims abstract description 56
- 238000005571 anion exchange chromatography Methods 0.000 title claims abstract description 45
- 238000002360 preparation method Methods 0.000 title claims description 31
- 229920000768 polyamine Polymers 0.000 claims abstract description 73
- 229920000642 polymer Polymers 0.000 claims abstract description 66
- 238000005349 anion exchange Methods 0.000 claims abstract description 12
- 239000000203 mixture Substances 0.000 claims abstract description 6
- 150000005846 sugar alcohols Polymers 0.000 claims abstract description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 33
- 150000003141 primary amines Chemical class 0.000 claims description 23
- 239000000741 silica gel Substances 0.000 claims description 23
- 229910002027 silica gel Inorganic materials 0.000 claims description 23
- IJKVHSBPTUYDLN-UHFFFAOYSA-N dihydroxy(oxo)silane Chemical compound O[Si](O)=O IJKVHSBPTUYDLN-UHFFFAOYSA-N 0.000 claims description 21
- 229920000333 poly(propyleneimine) Polymers 0.000 claims description 17
- 239000004593 Epoxy Substances 0.000 claims description 14
- 239000004005 microsphere Substances 0.000 claims description 13
- 150000001875 compounds Chemical class 0.000 claims description 8
- 238000004132 cross linking Methods 0.000 claims description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- LRWZZZWJMFNZIK-UHFFFAOYSA-N 2-chloro-3-methyloxirane Chemical compound CC1OC1Cl LRWZZZWJMFNZIK-UHFFFAOYSA-N 0.000 claims description 5
- 150000001412 amines Chemical class 0.000 claims description 5
- 238000001179 sorption measurement Methods 0.000 claims description 5
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000003431 cross linking reagent Substances 0.000 claims description 4
- 229910052710 silicon Inorganic materials 0.000 claims description 4
- 239000010703 silicon Substances 0.000 claims description 4
- MPPPKRYCTPRNTB-UHFFFAOYSA-N 1-bromobutane Chemical compound CCCCBr MPPPKRYCTPRNTB-UHFFFAOYSA-N 0.000 claims description 3
- VFWCMGCRMGJXDK-UHFFFAOYSA-N 1-chlorobutane Chemical compound CCCCCl VFWCMGCRMGJXDK-UHFFFAOYSA-N 0.000 claims description 3
- AOBIOSPNXBMOAT-UHFFFAOYSA-N 2-[2-(oxiran-2-ylmethoxy)ethoxymethyl]oxirane Chemical compound C1OC1COCCOCC1CO1 AOBIOSPNXBMOAT-UHFFFAOYSA-N 0.000 claims description 3
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 229920006037 cross link polymer Polymers 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 230000008859 change Effects 0.000 claims description 2
- 150000002081 enamines Chemical class 0.000 claims description 2
- GKIPXFAANLTWBM-UHFFFAOYSA-N epibromohydrin Chemical compound BrCC1CO1 GKIPXFAANLTWBM-UHFFFAOYSA-N 0.000 claims 1
- 230000015784 hyperosmotic salinity response Effects 0.000 abstract description 4
- 239000003957 anion exchange resin Substances 0.000 abstract description 2
- 125000002924 primary amino group Chemical class [H]N([H])* 0.000 abstract 2
- 238000000034 method Methods 0.000 description 23
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000007788 liquid Substances 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 239000011148 porous material Substances 0.000 description 7
- 238000001035 drying Methods 0.000 description 6
- 239000000377 silicon dioxide Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- -1 DEAE tertiary amine Chemical class 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 description 4
- 150000001450 anions Chemical class 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000002131 composite material Substances 0.000 description 4
- 238000002086 displacement chromatography Methods 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- 150000003335 secondary amines Chemical class 0.000 description 3
- CYNYIHKIEHGYOZ-UHFFFAOYSA-N 1-bromopropane Chemical compound CCCBr CYNYIHKIEHGYOZ-UHFFFAOYSA-N 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- 239000003463 adsorbent Substances 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 229960000074 biopharmaceutical Drugs 0.000 description 2
- 238000005253 cladding Methods 0.000 description 2
- 239000000084 colloidal system Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000007306 functionalization reaction Methods 0.000 description 2
- 238000005470 impregnation Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229910021426 porous silicon Inorganic materials 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000003980 solgel method Methods 0.000 description 2
- 238000000527 sonication Methods 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000011210 chromatographic step Methods 0.000 description 1
- 239000012539 chromatography resin Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000011026 diafiltration Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000012444 downstream purification process Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/26—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
- B01J13/06—Making microcapsules or microballoons by phase separation
- B01J13/14—Polymerisation; cross-linking
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2201/00—Foams characterised by the foaming process
- C08J2201/04—Foams characterised by the foaming process characterised by the elimination of a liquid or solid component, e.g. precipitation, leaching out, evaporation
- C08J2201/044—Elimination of an inorganic solid phase
- C08J2201/0442—Elimination of an inorganic solid phase the inorganic phase being a metal, its oxide or hydroxide
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2339/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a single or double bond to nitrogen or by a heterocyclic ring containing nitrogen; Derivatives of such polymers
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2339/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a single or double bond to nitrogen or by a heterocyclic ring containing nitrogen; Derivatives of such polymers
- C08J2339/02—Homopolymers or copolymers of vinylamine
Landscapes
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Dispersion Chemistry (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The present invention provides a kind of salt tolerant anion-exchange chromatography medium, the salt tolerant anion-exchange chromatography medium is Cross-linked polyamines polyalcohol stephanoporate microballoons；The skeleton main structure of the salt tolerant anion-exchange chromatography medium is polyamines polymer, and highdensity primary amine aglucon is had on the polyamines polymer.Since the intrinsic composition of cross linked porous polyamines polymer microballoon of the invention is polyamines polymer, the anion exchange resin in its surface primary amine group density ratio existing product is high.Therefore it not only has salt tolerance anion exchange absorbing function, but also adsorbing carrying capacity can be higher.
Description
Technical field
The invention belongs to chromatography media fields, and in particular to a kind of salt tolerant anion-exchange chromatography medium.
Background technique
Designed for bio-pharmaceuticals drug production most of downstream purification process in, dilution and diafiltration sequence be can not
The previous work avoided.These operations are commonly used in optimum condition needed for feed liquid is adjusted to optimised process performance.However, this
A little steps usually take considerable time, water and labour.Since bio-pharmaceuticals produces the driving reduced more and more by cost, because
This improve process economy one possible way to be by as far as possible before chromatographic step or between eliminate these units behaviour
Make to simplify purifying.
In the biology manufacture purifying process such as many therapeutic recombinant proteins, nucleic acid or virion, anion exchange
Adsorbent is frequently used for the capture purification step of the first step.However, being needed using conventional anion displacement chromatography medium lower
Ionic strength is to obtain enough carrying capacity.It is thus typically necessary to by feed liquid dilute or be percolated so that feed conductivity be reduced to it is suitable
Close conventional anion adsorbent.Use " salt tolerant " (salt-tolerant) anion exchange absorbing then can be directly from undiluted
Raw material in capture and significantly improve process economy.In addition, in many antibody biologies manufacture purifying process, anion exchange
It is then frequently used for after Protein A capture, or flows through mode removal of impurities (DNA, disease after cationic consummate (polish)
Poison, HCP, polymer etc.).Under both of these case feed liquid often contain in until high salt concentration, conventional anion chromatographic resin exists
Under this high conductivity almost can not adsorption-edulcoration, therefore be usually also required to by feed liquid dilute or be percolated so that feed conductivity drop
It is low.Feed liquid can not be diluted if using " salt tolerant " anion exchange absorbing and directly flows through removal of impurities, improve process economy.
Anion chromatography medium with " salt tolerant " effect usually contains highdensity primary amine or secondary amine aglucon.Conventional Q
The strong negative or DEAE tertiary amine weak anionic displacement chromatography resin of quaternary amine is when conductivity is greater than 18s/cm almost without ion exchange
Absorption, and the chromatography media containing primary amine or secondary amine aglucon can be realized under higher salt concentrations (conductivity be greater than 20s/cm) from
Sub- exchange adsorption, this may be because primary amine or secondary amine can also be had an effect other than charge effect by hydrogen bond and protein
(bibliography：J Chromatogr A.1016(1)：21-33).Salt tolerant anion-exchange chromatography medium is all in the prior art
The polyamines polymer such as the polyvinylamine by key and with high density primary amine or polypropylene amine is prepared to stationary phase.
United States Patent (USP) US 5304638, which has been reported, is grafted to agarose gel chromatography dielectric surface preparation yin for polypropylene amine
Ion-exchange chromatography media.
United States Patent (USP) US 20110065900A1 and " J.Chromatogr.A 1305 (2013), 85-93 " reported table
Face has been bonded Sepharose FF chromatographic resin (0.25M chlorination under high salt conditions than conventional Q Sepharose of polypropylene amine
Sodium) anion exchange absorbing carrying capacity is significantly high.
United States Patent (USP) US8435406 reported by polypropylene amine cladding (coating) prepared in chromatographic film salt tolerant yin from
Sub- displacement chromatography film.
U.S. Patent application US 20140336355A1 has reported bonding and cross-linked polyvinylamine is solid in polystyrene microsphere
Fixed resistance to anion-exchange chromatography medium with high salt mutually prepared above.
The above prior art is all to be bonded or be coated on fixation for polyamines polymer (polyvinylamine or polypropylene amine)
Salt tolerant anion-exchange chromatography medium is prepared in phase.Product prepared by such preparation method can have a disadvantage that：1) its
Preparation method is all by polyamines polymer-bound or to be coated on chromatography media, and the primary amine quantity of introducing is limited；2) lead to
Polyamines polymer is introduced into chromatography media duct by the method for crossing bonding or cladding, and chromatography can be reduced during introducing
The aperture of medium and carrying capacity, or even since uncontrollability will lead to part chromatography media duct obstruction.
Summary of the invention
In order to solve the above technical problems, the invention discloses a kind of salt tolerant anion-exchange chromatography medium and its preparation sides
Method, which not only has salt tolerance anion exchange absorbing function, but also it is higher to adsorb carrying capacity.
To realize the above-mentioned technical purpose, the technical scheme is that：A kind of salt tolerant anion-exchange chromatography medium, it is described
Salt tolerant anion-exchange chromatography medium is Cross-linked polyamines polyalcohol stephanoporate microballoons；The salt tolerant anion-exchange chromatography medium
Skeleton main structure is polyamines polymer, and primary amine aglucon is had on the polyamines polymer.
Wherein, the salt tolerant anion-exchange chromatography medium is Cross-linked polyamines polyalcohol stephanoporate microballoons, is referred to described
Polyamines polymer is fixed on chromatography media in a manner of being crosslinked.
Wherein, highdensity primary amine aglucon is had on the polyamines polymer.
Wherein, the partial size of the salt tolerant anion-exchange chromatography medium is 5-100um；The salt tolerant anion-exchange chromatography
Medium is porous media, and aperture is
Preferably, the salt tolerant anion-exchange chromatography medium is porous media, and aperture is
Wherein, the polyamines polymer is polyvinylamine and/or polypropylene amine.
The present invention also provides a kind of preparation method of salt tolerant anion-exchange chromatography medium, the preparation method includes following
Step：It 1) will be in the hole of polyamines Polymer adsorption to Bio-sil；2) polyamines crosslinked polymer is fixed on by introducing crosslinked agent
Inside porous silicon ball, Cross-linked polyamines polymer/silica gel complex microsphere is formed；3) by the Cross-linked polyamines polymer/silica gel
Bio-sil in complex microsphere is dissolved by highly basic, forms skeleton main structure as band on polyamines polymer, polyamines polymer
There is the salt tolerant anion-exchange chromatography medium of primary amine aglucon.
Wherein, polyamines polymer molecular weight is greater than 1000 in step 1).
Preferably, polyamines polymer molecular weight is greater than 10000 in step 1).
Wherein, the polyamines polymer is polyvinylamine and/or polypropylene amine.
Wherein, crosslinking agent described in step 2) is difunctional or multi-group epoxy compound.
Preferably, the difunctional or multi-group epoxy compound include epoxychloropropane, epoxy chlorobutane, epoxy
N-Propyl Bromide, epoxy bromobutane, butyl bisglycidyl ether, any or mixture in ethylene glycol bisglycidyl ether.
Wherein, highdensity primary amine aglucon is had on the polyamines polymer.
The polyamines polymer is the high-molecular compound with high density primary amine.
The present invention is template with Bio-sil microballoon, and the silicone hydroxyl on Bio-sil surface has negative electrical charge, polyamines polymer
It is positively charged, therefore inside Bio-sil microballoon easily adsorbed polyamines polymer to its duct.Meanwhile the present invention uses excessively
Polyamines polymer, be filled the duct of Bio-sil microballoon completely.A small amount of small molecule crosslinking agent is added will be more in duct
Amine polymer crosslinking is fixed on inside the duct of Bio-sil microballoon, forms Cross-linked polyamines polymer/silica gel complex microsphere.So
Afterwards, the processing of this complex microsphere sodium hydroxide strong base solution is dissolved into Bio-sil template ingredient, obtained cross linked porous more
Amine polymer microballoon.Since the intrinsic composition of cross linked porous polyamines polymer microballoon of the invention is polyamines polymer, this
Invention can not only provide the primary amine aglucon of salt tolerant, and the primary amine group density of microsphere surface is also than the anion exchange in existing product
Resin is much higher, is very suitable to carry out enrichment use under high salt concn.
Polyamines polymer is introduced into chromatography media duct by the present invention in the method being crosslinked, so as to introduce high density
Primary amine quantity；Also, when cross-linking method introduces polyamines polymer, aperture and the carrying capacity of chromatography media will not be reduced, will not be hindered
Consent road.Therefore, salt tolerant anion-exchange chromatography medium of the invention not only has salt tolerance anion exchange absorbing function, but also
It is higher to adsorb carrying capacity.
The preparation method of salt tolerant chromatography media of the present invention is to pass through the silica gel conduct with porous structure using template
Template can select the silica gel of different pore size as template, the polyamines of different pore size and partial size can be obtained during the preparation process
Polymer chromatography medium.And the aperture of chromatography media and the molecular size of material to be separated are closely bound up, by the method for the invention may be used
To prepare the chromatography media of a variety of different pore sizes, partial size, the substance suitable for a variety of different molecular weight sizes is separated.
Detailed description of the invention
Fig. 1 is the step schematic diagram that salt tolerant anion-exchange chromatography medium is prepared using the method for the present invention.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer with reference to specific embodiments to this
The technical solution of invention further illustrates.It should be understood that these embodiments are merely to illustrate the present invention rather than the limitation present invention
Range.In addition, it should also be understood that, after reading the contents of the present invention, those skilled in the art can make various change to the present invention
Dynamic or modification, such equivalent forms equally fall within limited range of the present invention.
A kind of salt tolerant anion-exchange chromatography medium, the salt tolerant anion-exchange chromatography medium are the polymerizations of Cross-linked polyamines
Object porous microsphere；The skeleton main structure of the salt tolerant anion-exchange chromatography medium is polyamines polymer, poly- in the polyamines
It closes and has primary amine aglucon on object.
Wherein, highdensity primary amine aglucon is had on the polyamines polymer.
Wherein, the partial size of the salt tolerant anion-exchange chromatography medium is 5-100um；The salt tolerant anion-exchange chromatography
Medium is porous media, and aperture is
Wherein, the polyamines polymer is polyvinylamine and/or polypropylene amine.
The present invention also provides a kind of preparation method of salt tolerant anion-exchange chromatography medium, the preparation method includes following
Step：It 1) will be in the hole of polyamines Polymer adsorption to Bio-sil；2) polyamines crosslinked polymer is fixed on by introducing crosslinked agent
Inside porous silicon ball, Cross-linked polyamines polymer/silica gel complex microsphere is formed；3) by the Cross-linked polyamines polymer/silica gel
Bio-sil in complex microsphere is dissolved by highly basic, forms skeleton main structure as band on polyamines polymer, polyamines polymer
There is the salt tolerant anion-exchange chromatography medium of primary amine aglucon.
Wherein, polyamines polymer molecular weight is greater than 1000 in step 1).
Preferably, polyamines polymer molecular weight is greater than 10000 in step 1).
Wherein, the polyamines polymer is polyvinylamine and/or polypropylene amine.
Wherein, crosslinking agent described in step 2) is difunctional or multi-group epoxy compound.
Preferably, the difunctional or multi-group epoxy compound include epoxychloropropane, epoxy chlorobutane, epoxy
N-Propyl Bromide, epoxy bromobutane, butyl bisglycidyl ether, any or mixture in ethylene glycol bisglycidyl ether.
Wherein, highdensity primary amine aglucon is had on the polyamines polymer.
The polyamines polymer is the high-molecular compound with high density primary amine.
Embodiment 1
Preparation process：
Purchase voluntarily prepares Bio-sil template.Preparation method generally with ethyl orthosilicate (TEOS) be silicon source,
Can be used spray drying process, polymerisation induced colloid aggregation method (stacked silica bead method), sol-gel method (can refer to Unger K,
Schick-Kalb J, Krebs K F.Preparation of porous silica spheres for column
Liquid [J] .Journal of Chromatography A, 1973,83 (AUG29)：And template (Jiang Biwang, Wu Jun 5.)
At equal grain porous silica microballoon of Chen Rong Ji's functionalization and its preparation method and application：, CN102070152A [P]
The methods of) .2011. preparation.
Step 1：Polyvinylamine is adsorbed onto the hole of ready Bio-sil
Weigh 10g Bio-sil (partial size 10um, apertureSurface area is 300m2/ g, pore volume 0.8cm3/ g),
6h is dried under vacuum condition.The silica gel sample handled well is added to the polyvinylamine (molecular weight that mass fraction is 60%
For thorough impregnation in 20000) aqueous solution, 12h is mixed, after stir drying under the conditions of 60 DEG C, it is rear that move into 65 DEG C of vacuum dry
In dry case until weight no longer changes, silica gel/polyvinylamine solid composite is obtained.
Step 2：Cross-linked polyvinylamine
Silica gel after above-mentioned drying/polyvinylamine solid composite is distributed in isopropanol, 10ml is added after stirring 1h
Epoxychloropropane heats up 60 DEG C and carries out cross-linking reaction.It filters, washing, is moved into 65 DEG C of vacuum ovens after the completion of cross-linking reaction
Drying obtains Cross-linked polyvinylamine/silica gel complex microsphere until weight no longer changes.
Step 3：Template removal
Obtained Cross-linked polyvinylamine/silica gel compound is distributed in 500ml 1.0M NaOH solution, is surpassed at room temperature
Sonication 60min, processing dissolves silica gel pattern mould.
Fig. 1 is the step schematic diagram that the method for the present invention prepares salt tolerant anion-exchange chromatography medium crosslinked poly amine.This hair
It is bright using monodispersed Bio-sil microballoon as template, by assembling polyamines polymer in duct, to form Cross-linked polyamines poly-
Object/silica gel complex microsphere is closed, then Bio-sil template is fallen by alkali liquid corrosion, obtains Cross-linked polyamines polyalcohol stephanoporate microballoons,
Precisely prepare polyamines polymer anion displacement chromatography medium.
Embodiment 2
Preparation process：
Purchase voluntarily prepares Bio-sil template.Preparation method generally with ethyl orthosilicate (TEOS) be silicon source,
Can be used spray drying process, polymerisation induced colloid aggregation method (stacked silica bead method), sol-gel method (can refer to Unger K,
Schick-Kalb J, Krebs K F.Preparation of porous silica spheres for column
Liquid [J] .Journal of Chromatography A, 1973,83 (AUG29)：And template (Jiang Biwang, Wu Jun 5.)
At equal grain porous silica microballoon of Chen Rong Ji's functionalization and its preparation method and application：, CN102070152A [P]
The methods of) .2011. preparation.
Step 1：Polypropylene amine is adsorbed onto the hole of ready Bio-sil
Weigh 10g Bio-sil (partial size 20um, aperture), 6h is dried under vacuum condition.It will handle well
Silica gel sample be added to thorough impregnation in polypropylene amine (molecular weight 20000) aqueous solution that mass fraction is 60%, mixing stirs
Mix 12h, after stir drying under the conditions of 60 DEG C, it is rear to move into 65 DEG C of vacuum ovens until weight no longer changes, obtain silicon
Glue/polypropylene amine solid composite.
Step 2：Crosslinked polypropylene enamine
Silica gel after above-mentioned drying/polypropylene amine solid composite is distributed in isopropanol, 10ml is added after stirring 1h
Epoxychloropropane heats up 60 DEG C and carries out cross-linking reaction.It filters, washing, is moved into 65 DEG C of vacuum ovens after the completion of cross-linking reaction
Drying obtains Cross-linked polypropylene amine/silica gel complex microsphere until weight no longer changes.
Step 3：Template removal
Obtained Cross-linked polypropylene amine/silica gel compound is distributed in 500ml 1.0M NaOH solution, is surpassed at room temperature
Sonication 60min, processing dissolve silica gel pattern mould.
The present invention using the polyamines polymer that is crosslinked be substrate formed microballoon as salt tolerant anion-exchange chromatography medium, it
Anion exchange resin in the primary amine group density ratio existing product on surface is high, therefore it not only has salt tolerance anion exchange
Adsorption function, and adsorbing carrying capacity can be higher.Polyamines polymer itself is both that medium stationary phase skeleton and resistance to salt anionic are handed over
Base is changed, therefore, the quantity of the anion exchange function group of dielectric surface i.e. highest, i.e., its salt resistant character is optimal
More.
The present invention uses excessive polyamines polymer, is filled the duct of Bio-sil microballoon completely, introduces enough
Primary amine quantity.Polyamines polymer is introduced into chromatography media duct by the method being crosslinked, chromatography media will not be reduced
Aperture and discretion.
In addition, the preparation method of this chromatography media is prepared by template, i.e., made by the silica gel with porous structure
For template, during the preparation process, the silica gel of different pore size can be selected as template, the more of different pore size and partial size can be obtained
Amine polymer chromatography media.And the aperture of chromatography media and the molecular size of material to be separated are closely bound up, can be made by this law
The chromatography media of standby different pore sizes a variety of out, partial size, the substance suitable for a variety of different molecular weight sizes separate.
Than the above described, the present invention can also have other modes realization, in the premise for not departing from the content of present invention
Under, it is any obviously replace it is within the scope of the present invention.
Claims (10)
1. a kind of salt tolerant anion-exchange chromatography medium, which is characterized in that the salt tolerant anion-exchange chromatography medium is crosslinking
Change polyamines polyalcohol stephanoporate microballoons；The skeleton main structure of the salt tolerant anion-exchange chromatography medium is polyamines polymer,
Primary amine aglucon is had on the polyamines polymer.
2. salt tolerant anion-exchange chromatography medium as described in claim 1, which is characterized in that the band on the polyamines polymer
There is highdensity primary amine aglucon.
3. salt tolerant anion-exchange chromatography medium as described in claim 1, which is characterized in that the salt tolerant anion exchange layer
The partial size for analysing medium is 5-100um；The salt tolerant anion-exchange chromatography medium is porous media, and aperture is
4. salt tolerant anion-exchange chromatography medium as described in claim 1, which is characterized in that the polyamines polymer is poly- second
Enamine and/or polypropylene amine.
5. a kind of preparation method of salt tolerant anion-exchange chromatography medium, which is characterized in that the preparation method includes following step
Suddenly：It 1) will be in the hole of polyamines Polymer adsorption to Bio-sil；2) polyamines crosslinked polymer is fixed on more by introducing crosslinked agent
Inside the silicon ball of hole, Cross-linked polyamines polymer/silica gel complex microsphere is formed；3) the Cross-linked polyamines polymer/silica gel is multiple
The Bio-sil closed in microballoon is dissolved by highly basic, forms skeleton main structure to have on polyamines polymer, polyamines polymer
The salt tolerant anion-exchange chromatography medium of primary amine aglucon.
6. the preparation method of salt tolerant anion-exchange chromatography medium as claimed in claim 5, which is characterized in that more in step 1)
The molecular weight of amine polymer is greater than 1000.
7. the preparation method of salt tolerant anion-exchange chromatography medium as claimed in claim 5, which is characterized in that more in step 1)
The molecular weight of amine polymer is greater than 10000.
8. the preparation method of salt tolerant anion-exchange chromatography medium as claimed in claim 5, which is characterized in that the polyamines is poly-
Closing object is polyvinylamine and/or polypropylene amine.
9. the preparation method of salt tolerant anion-exchange chromatography medium as claimed in claim 5, which is characterized in that institute in step 2)
Stating crosslinking agent is difunctional or multi-group epoxy compound.
10. the preparation method of salt tolerant anion-exchange chromatography medium as claimed in claim 9, which is characterized in that double officials
It can roll into a ball or multi-group epoxy compound includes epoxychloropropane, epoxy chlorobutane, epoxy bromopropane, epoxy bromobutane, butyl
Any or mixture in bisglycidyl ether, ethylene glycol bisglycidyl ether.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810963586.XA CN108892803B (en) | 2018-08-22 | 2018-08-22 | Salt-resistant anion exchange chromatography medium and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810963586.XA CN108892803B (en) | 2018-08-22 | 2018-08-22 | Salt-resistant anion exchange chromatography medium and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108892803A true CN108892803A (en) | 2018-11-27 |
| CN108892803B CN108892803B (en) | 2021-03-12 |
Family
ID=64358274
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810963586.XA Active CN108892803B (en) | 2018-08-22 | 2018-08-22 | Salt-resistant anion exchange chromatography medium and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108892803B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114989359A (en) * | 2022-06-22 | 2022-09-02 | 苏州纳微科技股份有限公司 | Cation exchange chromatography medium and preparation method thereof |
| WO2023012251A1 (en) * | 2021-08-05 | 2023-02-09 | Instraction Gmbh | Removal of viruses from water by filtration |
| CN117358321A (en) * | 2023-12-04 | 2024-01-09 | 赛普(杭州)过滤科技有限公司 | Chromatography medium and preparation method thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6151006A (en) * | 1984-08-20 | 1986-03-13 | Mitsubishi Chem Ind Ltd | Crosslinking ball making method for polyvinylamines |
| CN1760219A (en) * | 2005-10-21 | 2006-04-19 | 天津大学 | Method for preparing microspheres of rigid, cross linked porous polyacrylamide |
| CN102452663A (en) * | 2010-10-15 | 2012-05-16 | 中国石油化工股份有限公司 | Improved preparation method of EU-1 molecular sieve |
| CN102500300A (en) * | 2011-11-23 | 2012-06-20 | 浙江大学 | Method for preparing polymer porous microspheres through mixed phase separation |
| US20150344520A1 (en) * | 2014-05-27 | 2015-12-03 | Jnc Corporation | Chromatography media and protein purification method using the same |
| CN105531241A (en) * | 2013-09-20 | 2016-04-27 | 赫罗伊斯石英玻璃股份有限两合公司 | Method for producing porous carbon products |
| CN106025279A (en) * | 2016-07-01 | 2016-10-12 | 东莞市凯金新能源科技股份有限公司 | High-capacity porous spherical graphitized carbon negative electrode material and preparation method thereof |
-
2018
- 2018-08-22 CN CN201810963586.XA patent/CN108892803B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6151006A (en) * | 1984-08-20 | 1986-03-13 | Mitsubishi Chem Ind Ltd | Crosslinking ball making method for polyvinylamines |
| CN1760219A (en) * | 2005-10-21 | 2006-04-19 | 天津大学 | Method for preparing microspheres of rigid, cross linked porous polyacrylamide |
| CN102452663A (en) * | 2010-10-15 | 2012-05-16 | 中国石油化工股份有限公司 | Improved preparation method of EU-1 molecular sieve |
| CN102500300A (en) * | 2011-11-23 | 2012-06-20 | 浙江大学 | Method for preparing polymer porous microspheres through mixed phase separation |
| CN105531241A (en) * | 2013-09-20 | 2016-04-27 | 赫罗伊斯石英玻璃股份有限两合公司 | Method for producing porous carbon products |
| US20150344520A1 (en) * | 2014-05-27 | 2015-12-03 | Jnc Corporation | Chromatography media and protein purification method using the same |
| CN106025279A (en) * | 2016-07-01 | 2016-10-12 | 东莞市凯金新能源科技股份有限公司 | High-capacity porous spherical graphitized carbon negative electrode material and preparation method thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023012251A1 (en) * | 2021-08-05 | 2023-02-09 | Instraction Gmbh | Removal of viruses from water by filtration |
| CN114989359A (en) * | 2022-06-22 | 2022-09-02 | 苏州纳微科技股份有限公司 | Cation exchange chromatography medium and preparation method thereof |
| CN114989359B (en) * | 2022-06-22 | 2024-03-12 | 苏州纳微科技股份有限公司 | Cation exchange chromatography medium and preparation method thereof |
| CN117358321A (en) * | 2023-12-04 | 2024-01-09 | 赛普(杭州)过滤科技有限公司 | Chromatography medium and preparation method thereof |
| CN117358321B (en) * | 2023-12-04 | 2024-03-19 | 赛普(杭州)过滤科技有限公司 | Chromatography medium and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108892803B (en) | 2021-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Cai et al. | Application of deep eutectic solvents in chromatography: A review | |
| Sýkora et al. | Recent advances in mixed‐mode chromatographic stationary phases | |
| Li et al. | Affinity monolith chromatography: A review of general principles and applications | |
| CN102015094B (en) | Chromatography medium | |
| CN101939094B (en) | Method for production of separation media | |
| JPH06504482A (en) | Hydrophilic sulfonamide bond coating | |
| CN104945637B (en) | A kind of graft type retains by force the preparation method of anion chromatographic filling material | |
| CN108892803A (en) | A kind of salt tolerant anion-exchange chromatography medium and preparation method thereof | |
| BRPI0519950B1 (en) | chromatographic medium, column for chromatography and process for separating components from a solution | |
| JP6702587B2 (en) | Separation matrix for purification of biological particles | |
| Saylan et al. | Surface imprinting approach for preparing specific adsorbent for IgG separation | |
| CN102416313B (en) | Bisphenol A dummy template molecularly imprinted stir bar and preparation method thereof | |
| CN105131181B (en) | A kind of preparation method of ionic liquid hybrid imprinting material | |
| CN111659356B (en) | Preparation and application of a polyethyleneimine-modified reversed-phase/strong anion-exchange mixed-mode polymer | |
| US8945389B2 (en) | Method for substance separation using a cellulose hydrate membrane in size exclusion chromatography | |
| CN107722178A (en) | A kind of preparation method and application of the hollow porous type molecularly imprinted polymer of macrolide antibiotics | |
| CN104289209B (en) | A WCX/HIC bifunctional mixed-mode polymer matrix chromatographic stationary phase for protein separation and its preparation method | |
| CN108927010A (en) | Adsorption function membrane material and its preparation method and application | |
| CN106496483B (en) | A kind of magnetism hydrophilic molecules trace resin, preparation method and its application in aqueous medium sample | |
| CN104437438B (en) | Surface modification of resin chromatograph packing material and preparation method thereof and solid-phase extraction column | |
| CN116899633B (en) | Hydrophilic anion exchange chromatography medium and preparation method and application thereof | |
| Xu et al. | Establishment of a novel surface-imprinting system for melamine recognition and mechanism of template–matrix interactions | |
| CN111393705B (en) | Hydrophilic molecularly imprinted resin and preparation method and application thereof | |
| JPH11183460A (en) | Packing material for liquid chromatography and its production and use | |
| Liu et al. | Construction of amine‐rich coating utilizing amine curing reaction of triglycidyl isocyanate with triethylenetetramine on the surface of poly (styrene‐divinylbenzene) microspheres for preparation of anion exchange stationary phase |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |